Here, @JoshCofsky, @doudna_lab, and colleagues, use #CryoEM and biochemical analysis to investigate how the #CRISPR associated protein #Cas9 interrogates DNA to locate its RNA-matching target sequence @berkeleyMCB@igisci https://t.co/4xu21kF6bg
@KatieBerryRNA@dsashital@doudna_lab@gavinjknott I am SO grateful that MR worked—I was really dreading the prospect of hexammines and I know my own attempt would also have been disastrous 😂
Have you ever wondered how A-form and B-form nucleic acid helices might interact with each other during the expansion of a Cas9-bound R-loop? Check out our crystal structure of an interhelical stack in @PLOSONE ! #stacked
🙏 @doudna_lab@gavinjknott +CLG
https://t.co/4fupkhQVD4
"A tweak and a peek"! Our Cas9 paper is now out in @NatureSMB (https://t.co/8MoKSdmeMM). Thanks also to Sinan + Russell for the News & Views article (https://t.co/39RD3si49l), which nailed it with the title and the beautiful explanations :)
🙏@doudna_lab@gavinjknott@NogalesLab
1. I am thrilled to share the accepted version of our Cas9 mismatch manuscript, now in @Nature! Here’s a (not-so) brief outline of some of the findings from our original preprint, and some VERY exciting new findings https://t.co/etRmAw6WPW
@gavinjknott@SnowMedical@em6wong @Stephintje Incredible!!! But also totally credible because you rock Gavin!! Congrats to you and the Knott lab, present and future. So proud 🥂
The first crystal structure I solved myself, no thanks to some pesky diffraction anisotropy! With crucial help from @gavinjknott, Christine Gee, and @doudna_lab, of course. If you don't care about nucleic acid structure, just check out the sexy lattice architecture in Fig. 1 😍
Keeping the ball rolling with my next manuscript from the @MartinJinek lab! We solved several
structures of intermediary binding states of Cas9 to elucidate the mechanism of target DNA binding and conformational activation!
https://t.co/DMrfLCCI0a
(1/2) Cas9 can cut off-targets. We used kinetics-guided structural studies of Cas9 bound to different mismatches and reveal DNA bending requirements for targeting, the non-target strand poised for cleavage in the active site of RuvC, and… https://t.co/3ok1bCNBfG
New pre-print! While #CRISPR-Cas9 searches through the genome, how does it actually "read" DNA sequence information, which is normally hidden within the double helix? We determined structures of Cas9 in the act of DNA interrogation to find out! Thread below 👇 (1/8)
@BKleinstiver Thanks Ben! I'm getting a whole lot of questions along the lines of "but does SpRY Cas9 float around doing this at EVERY base pair?" Lots to think about!
@RossWilsonLab@pdhsu@gavinjknott@doudna_lab Yeah, it'll be interesting to get numbers on it! We expect the net ΔG for Cas9:DNA's bending transition to be much less negative (or maybe even positive?) than for other DNA-bending proteins because of Cas9's functional requirement to release off-target DNA ASAP